In Silico and In Vitro evaluation of newly synthesized Thiourea derivatives as potential chemopreventive agents against MSF-7 cancer cell line [EMBARGOED]

Abstract

The increase of cancer cases is correlated to the less selective, less effective, and toxicity of the available anticancer drugs, demanding for the development of new effective therapeutic agents. Epidermal growth factor receptor tyrosine kinase (EGFR-TK) is the critical mediator involving cell signaling pathways, has emerged as a promising target for pharmacological inhibition. Thiourea derivatives are widely explored as anticancer agents due to the presence of heteroatom groups, C=S (hydrogen bond acceptor) and NH (hydrogen bond donor), which are essential to enhance drug potency, target selectivity, provide structural diversity, and contribute to favourable drug metabolism and pharmacokinetic characteristics. In this study, a total of 80 thiourea derivatives were designed and screened for their absorption, distribution, metabolism, excretion and toxicity (ADMET) using Chemsketch 1.2, SwissADME and ADMETlab, respectively. All compounds showed satisfactory ADMET properties except for nitro bearing thiourea group. Subsequently, the molecular docking study was carried out via AutoDock Vina version 1.1.2 to identify the potential of designed compounds as EGFR TK inhibitor by determining their binding affinities and interaction patterns between the designed compounds and EGFR-TK binding site (PDB ID: 1M17). The docking results indicate that all thiourea derivatives have lower binding affinities (-7.3 to -8.7 kcal/mol) compared to standard drug erlotinib (-6.7 kcal/mol). The best compounds with lower binding affinities, compounds 1A, 1B, 1C and 1E were further analysed for their complexes stability through molecular dynamic (MD) simulation method. The results revealed that complex 1E, showed adequate compactness, uniqueness, and satisfactory stability. In an effort to discover the potential of the compounds as anticancer agents, synthesizing, characterizing and in vitro assays were applied. Compounds 1B, 1C and 1E were synthesized and characterized via CHNS elemental analysis, FTIR, 1H and 13C NMR. Micro elemental analysis data of the compounds were in agreement with the theoretical values. In IR analysis, all synthesized compounds depicted important absorption bands such as ѵ(NH), ѵ(C=ONH) and ѵ(C=S). For NMR analyses, the resonance of important chemical shifts can be observed at expected regions NH (δH:10.86-11.69 ppm) in 1H NMR and, C=O (δC: 156.55-167.29 ppm) and C=S (δC: 171.32-181.48 ppm) in 13C NMR. The MTT assay technique against human breast adenocarcinoma cell line (MCF-7) were employed to evaluate the cytotoxicity effects of these compounds. The results revealed that all synthesized compounds have shown good cytotoxic activity with IC50 value 21.91 µM (1E), 29.66 µM (1C) and 44.19 µM (1B) compared to the previous reported erlotinib with IC50: 87.34 µM. In a nutshell, this study suggested that the designed thiourea derivatives based on crucial EGFR-TKI pharmacophoric features can be a promising candidate for EGFR-TKI, which could be useful in designing more potent and drug-like EGFR-TKIs in the future.

The increase of cancer cases is correlated to the less selective, less effective, and toxicity of the available anticancer drugs, demanding for the development of new effective therapeutic agents. Epidermal growth factor receptor tyrosine kinase (EGFR-TK) is the critical mediator involving cell signaling pathways, has emerged as a promising target for pharmacological inhibition. Thiourea derivatives are widely explored as anticancer agents due to the presence of heteroatom groups, C=S (hydrogen bond acceptor) and NH (hydrogen bond donor), which are essential to enhance drug potency, target selectivity, provide structural diversity, and contribute to favourable drug metabolism and pharmacokinetic characteristics. In this study, a total of 80 thiourea derivatives were designed and screened for their absorption, distribution, metabolism, excretion and toxicity (ADMET) using Chemsketch 1.2, SwissADME and ADMETlab, respectively. All compounds showed satisfactory ADMET properties except for nitro bearing thiourea group. Subsequently, the molecular docking study was carried out via AutoDock Vina version 1.1.2 to identify the potential of designed compounds as EGFR-TK inhibitor by determining their binding affinities and interaction patterns between the designed compounds and EGFR-TK binding site (PDB ID: 1M17). The docking results indicate that all thiourea derivatives have lower binding affinities (-7.3 to -8.7 kcal/mol) compared to standard drug erlotinib (-6.7 kcal/mol). The best compounds with lower binding affinities, compounds 1A, 1B, 1C and 1E were further analysed for their complexes stability through molecular dynamic (MD) simulation method. The results revealed that complex 1E, showed adequate compactness, uniqueness, and satisfactory stability. In an effort to discover the potential of the compounds as anticancer agents, synthesizing, characterizing and in vitro assays were applied. Compounds 1B, 1C and 1E were synthesized and characterized via CHNS elemental analysis, FTIR, 1H and 13C NMR. Micro elemental analysis data of the compounds were in agreement with the theoretical values. In IR analysis, all synthesized compounds depicted important absorption bands such as ?(NH), ?(C=ONH) and ?(C=S). For NMR analyses, the resonance of important chemical shifts can be observed at expected regions NH (?H:10.86-11.69 ppm) in 1H NMR and, C=O (?C: 156.55-167.29 ppm) and C=S (?C: 171.32-181.48 ppm) in 13C NMR. The MTT assay technique against human breast adenocarcinoma cell line (MCF-7) were employed to evaluate the cytotoxicity effects of these compounds. The results revealed that all synthesized compounds have shown good cytotoxic activity with IC50 value 21.91 µM (1E), 29.66 µM (1C) and 44.19 µM (1B) compared to the previous reported erlotinib with IC50: 87.34 µM. In a nutshell, this study suggested that the designed thiourea derivatives based on crucial EGFR-TKI pharmacophoric features can be a promising candidate for EGFR-TKI, which could be useful in designing more potent and drug-like EGFR-TKIs in the future.